骨形成蛋白cDNA的定位插入突变及其克隆

Directed mutagenesis and cloning of bone morphogenetic protein cDNA

用寡核苷酸诱导的定位突变技术,将骨形成蛋白基因5’端经突变后去除了信号肽序列并,引入限制性内切酶NcoI位点及起始密码ATG,将此片段重新组装后克隆入pBR322质粒中,为进一步使基因克隆和表达有活性的骨形成蛋白奠定了基础。

Using oligonucleotide directed mutagenesis techniques, the cDNA gene of human bone morphogenetic protein (hBMP^l) was designed for reforming to express in E. coli. The leading sequence located at 5'-end of the gene was deleted and translational starting codon with a restriction endonuclease Ncol site was introduced .The reformed gene was inserted into plasmid pBR322 and recombinant was transformed with E. coli HB101- Analysis with restriction enzyme mapping and DNA sequencing shows that the gene mutagenesis is successfully in according to the original designation.